当前位置: X-MOL 学术ACS Nano › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Nanometer-Resolved Mapping of Cell–Substrate Distances of Contracting Cardiomyocytes Using Surface Plasmon Resonance Microscopy
ACS Nano ( IF 15.8 ) Pub Date : 2018-09-04 00:00:00 , DOI: 10.1021/acsnano.8b01396
Eva Kreysing , Hossein Hassani , Nico Hampe , Andreas Offenhäusser

It has been shown that quantitative measurements of the cell–substrate distance of steady cells are possible with scanning surface plasmon resonance microscopy setups in combination with an angle resolved analysis. However, the accuracy of the determined cell–substrate distances as well as the capabilities for the investigation of cell dynamics remained limited due to the assumption of a homogeneous refractive index of the cytosol. Strong spatial or temporal deviations between the local refractive index and the average value can result in errors in the calculated cell–substrate distance of around 100 nm, while the average accuracy was determined to 37 nm. Here, we present a combination of acquisition and analysis techniques that enables the measurement of the cell–substrate distance of contractile cells as well as the study of intracellular processes through changes in the refractive index at the diffraction limit. By decoupling the measurement of the cell–substrate distance and the refractive index of the cytoplasm, we could increase the accuracy of the distance measurement on average by a factor of 25 reaching 1.5 nm under ideal conditions. We show a temporal and spatial mapping of changes in the refractive index and the cell–substrate distance which strongly correlate with the action potentials and reconstruct the three-dimensional profile of the basal cell membrane and its dynamics, while we reached an actual measurement accuracy of 2.3 nm.

中文翻译:

使用表面等离子体共振显微镜对收缩心肌细胞-基质距离的纳米分辨定位

研究表明,结合扫描表面等离子体共振显微镜设置和角度分辨分析,可以对稳定细胞的细胞-基质距离进行定量测量。但是,由于假定了细胞质的折射率均一,所以确定的细胞与底物之间的距离的准确性以及研究细胞动力学的能力仍然受到限制。局部折射率与平均值之间的强烈空间或时间偏差可能会导致计算出的单元-衬底距离大约为100 nm,而平均精度被确定为37 nm。这里,我们提出了一种采集和分析技术的组合,该技术能够测量可收缩细胞的细胞-底物距离,以及通过衍射极限处的折射率变化来研究细胞内过程。通过将细胞-基质距离的测量值与细胞质的折射率解耦,我们可以在理想条件下将距离测量的精度平均提高25倍,达到1.5 nm。我们显示了折射率和细胞-基质距离的变化的时间和空间映射,这些变化与动作电位密切相关,并重建了基底细胞膜的三维轮廓及其动力学,同时达到了实际的测量精度2.3纳米
更新日期:2018-09-04
down
wechat
bug